Song Yilan, Wang Zhiguang, Jiang Jingzhi, Piao Yihua, Li Li, Xu Chang, Piao Hongmei, Li Liangchang, Yan Guanghai
Jilin Key Laboratory for Immune and Targeting Research on Common Allergic Diseases, Yanbian University, Yanji, China.
Department of Anatomy, Histology and Embryology, Yanbian University Medical College, Yanji, China.
J Cell Mol Med. 2020 Dec;24(23):13739-13750. doi: 10.1111/jcmm.15942. Epub 2020 Oct 30.
This study is to investigate the inhibitory effects and mechanisms of DEK-targeting aptamer (DTA-64) on epithelial mesenchymaltransition (EMT)-mediated airway remodelling in mice and human bronchial epithelial cell line BEAS-2B. In the ovalbumin (OVA)-induced asthmatic mice, DTA-64 significantly reduced the infiltration of eosinophils and neutrophils in lung tissue, attenuated the airway resistance and the proliferation of goblet cells. In addition, DTA-64 reduced collagen deposition, transforming growth factor 1 (TGF-β1) level in BALF and IgE levels in serum, balanced Th1/Th2/Th17 ratio, and decreased mesenchymal proteins (vimentin and α-SMA), as well as weekend matrix metalloproteinases (MMP-2 and MMP-9) and NF-κB p65 activity. In the in vitro experiments, we used TGF-β1 to induce EMT in the human epithelial cell line BEAS-2B. DEK overexpression (ovDEK) or silencing (shDEK) up-regulated or down-regulated TGF-β1 expression, respectively, on the contrary, TGF-β1 exposure had no effect on DEK expression. Furthermore, ovDEK and TGF-β1 synergistically promoted EMT, whereas shDEK significantly reduced mesenchymal markers and increased epithelial markers, thus inhibiting EMT. Additionally, shDEK inhibited key proteins in TGF-β1-mediated signalling pathways, including Smad2/3, Smad4, p38 MAPK, ERK1/2, JNK and PI3K/AKT/mTOR. In conclusion, the effects of DTA-64 against EMT of asthmatic mice and BEAS-2B might partially be achieved through suppressing TGF-β1/Smad, MAPK and PI3K signalling pathways. DTA-64 may be a new therapeutic option for the management of airway remodelling in asthma patients.
本研究旨在探讨靶向DEK的适体（DTA-64）对小鼠和人支气管上皮细胞系BEAS-2B中上皮-间质转化（EMT）介导的气道重塑的抑制作用及机制。在卵清蛋白（OVA）诱导的哮喘小鼠中，DTA-64显著减少肺组织中嗜酸性粒细胞和中性粒细胞的浸润，减轻气道阻力和杯状细胞的增殖。此外，DTA-64减少胶原沉积、支气管肺泡灌洗液中转化生长因子1（TGF-β1）水平和血清中IgE水平，平衡Th1/Th2/Th17比例，并降低间充质蛋白（波形蛋白和α-平滑肌肌动蛋白）以及基质金属蛋白酶（MMP-2和MMP-9）和核因子κB p65活性。在体外实验中，我们用TGF-β1诱导人上皮细胞系BEAS-2B发生EMT。DEK过表达（ovDEK）或沉默（shDEK）分别上调或下调TGF-β1表达，相反，TGF-β1暴露对DEK表达无影响。此外，ovDEK和TGF-β1协同促进EMT，而shDEK显著降低间充质标志物并增加上皮标志物，从而抑制EMT。此外，shDEK抑制TGF-β1介导的信号通路中的关键蛋白，包括Smad2/3、Smad4、p38丝裂原活化蛋白激酶、细胞外信号调节激酶1/2、c-Jun氨基末端激酶和磷脂酰肌醇-3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白。总之，DTA-64对哮喘小鼠和BEAS-2B的EMT的作用可能部分是通过抑制TGF-β1/Smad、丝裂原活化蛋白激酶和磷脂酰肌醇-3-激酶信号通路实现的。DTA-64可能是哮喘患者气道重塑管理的一种新的治疗选择。
